Photodynamic Activity of Protoporphyrin IX-Immobilized Cellulose Monolith for Nerve Tissue Regeneration.

The development of nerve conduits with a three-dimensional porous structure has attracted great attention as they closely mimic the major features of the natural extracellular matrix of the nerve tissue. As low levels of reactive oxygen species (ROS) function as signaling molecules to promote cell proliferation and growth, this study aimed to fabricate protoporphyrin IX (PpIX)-immobilized cellulose (CEPP) monoliths as a means to both guide and stimulate nerve regeneration. CEPP monoliths can be fabricated via a simple thermally induced phase separation method and surface modification. The improved nerve tissue regeneration of CEPP monoliths was achieved by the activation of mitogen-activated protein kinases, such as extracellular signal-regulated kinases (ERKs). The resulting CEPP monoliths exhibited interconnected microporous structures and uniform morphology. The results of in vitro bioactivity assays demonstrated that the CEPP monoliths with under 0.54 ± 0.07 µmol/g PpIX exhibited enhanced photodynamic activity on Schwann cells via the generation of low levels of ROS. This photodynamic activation of the CEPP monoliths is a cell-safe process to stimulate cell proliferation without cytotoxic side effects. In addition, the protein expression of phospho-ERK increased considerably after the laser irradiation on the CEPP monoliths with low content of PpIX. Therefore, the CEPP monoliths have a potential application in nerve tissue regeneration as new nerve conduits.

In vitro responses of neurofibroma fibroblasts, mast cells and Schwann cells obtained from patients with neurofibromatosis 1 to 308-nm excimer light and/or vitamin D3.

Fibroblasts, mast cells and Schwann cells were isolated from neurofibromas of patients with neurofibromatosis 1, and their responses to 308-nm excimer light irradiation and/or vitamin D3 or an analog (tacalcitol; 1,24-dihydroxyvitamin D3 ) were examined in vitro. Excimer light irradiation (300 mJ/cm(2) ) suppressed the growth of all three cell types. Exposure to 10(-7)  mol/L of 1α,25(OH)2 D3 (VD3 ) or tacalcitol suppressed the growth of fibroblasts and mast cells, but not Schwann cells. These results suggest that the different neurofibroma cell types show different responses to VD3 . A combination of excimer light irradiation and VD3 is necessary to suppress the growth of neurofibroma cells in vivo.

Effects of low level laser therapy on proliferation and neurotrophic factor gene expression of human schwann cells in vitro.

Previous studies have been proposed that proliferation and release of certain growth factors by different types of cells can be modulated by low level laser therapy. We aimed to demonstrate the effect of laser irradiation on human schwann cell proliferation and neurotrophic factor gene expression in vitro. Human schwann cells (SCs) were harvested from sural nerve that was obtained from organ donor followed by treatment with an 810 nm, 50 mW diode laser (two different energies: 1 J/cm(2) and 4 J/cm(2)) in three consecutive days. SC proliferation was measured, after first irradiation on days 1, 4 and 7 by the MTT assay. Real time PCR analysis was utilized on days 5 and 20 to evaluate the expression of key genes involved in nerve regeneration consist of NGF, BDNF and GDNF. Evaluation of cellular proliferation following one day after laser treatment revealed significant decrease in cell proliferation compared to control group. However on day 7, significant increase in proliferation was found in both the irradiated groups in comparison with the control group. No significant difference was found between the laser treated groups. Treatment of SCs with laser resulted in significant increase in NGF gene expression on day 20. Difference between two treated groups and control group was not significant for BDNF and GDNF gene expression. Our results demonstrate that low level laser therapy stimulate human schwann cell proliferation and NGF gene expression in vitro.